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TMBS4 Medium (DSMZ Medium 559)

[所属分类:培养基配方] [发布时间:2021-9-16] [发布人:网站管理员2] [阅读次数:] [返回]

TMBS4 Medium

(DSMZ Medium 559)

山东拓普生物工程有限公司培养基配方 http://www.topbiol.com

Composition per 1003.0mL:

Solution A..............................................................................870.0mL

Solution C ..............................................................................100.0mL

Solution D................................................................................10.0mL

Solution E (Vitamin solution)..................................................10.0mL

Solution F.................................................................................10.0mL

Solution B (Trace elements solution SL-10) .............................1.0mL

Solution G..................................................................................1.0mL

Solution H..................................................................................1.0mL

pH 7.1–7.4 at 25°C

Solution A:

Composition per 870.0mL:

NaCl.............................................................................................. 1.0g

MgCl2·6H2O ................................................................................. 0.4g

Na2SO4.......................................................................................... 3.0g

KCl................................................................................................ 0.5g

NH4Cl ........................................................................................... 0.3g

KH2PO4......................................................................................... 0.2g

CaCl2·2H2O ................................................................................ 0.15g

Resazurin ................................................................................... 1.0mg

Preparation of Solution A: Add components to distilled/deionized

water and bring volume to 870.0mL Mix thoroughly.

Solution B (Trace Elements Solution SL-10):

Composition per liter:

FeCl2·4H2O................................................................................... 1.5g

CoCl2·6H2O ........................................................................... 190.0mg

MnCl2·4H2O .......................................................................... 100.0mg

ZnCl2........................................................................................ 70.0mg

Na2MoO4·2H2O ....................................................................... 36.0mg

NiCl2·6H2O.............................................................................. 24.0mg

H3BO3 ........................................................................................ 6.0mg

CuCl2·2H2O ............................................................................... 2.0mg

HCl (25% solution)..................................................................10.0mL

Preparation of Solution B (Trace Elements Solution SL-10):

Add FeCl2·4H2O to 10.0mL of HCl solution. Mix thoroughly. Add dis

tilled/deionized water and bring volume to 1.0L. Add remaining com

ponents. Mix thoroughly. Sparge with 100% N2. Autoclave for 15 min

at 15 psi pressure–121°C.

Solution C:

Composition per 100.0mL:

NaHCO3........................................................................................ 5.0g

Preparation of Solution C: Add NaHCO3 to distilled/deionized

water and bring volume to 100.0mL Mix thoroughly. Filter sterilize.

Flush with 80% N2 + 20% CO2 to remove dissolved oxygen.

Solution D:

Composition per 10.0mL:

Syringic acid ................................................................................. 0.6g

Preparation of Solution D: Add syringic acid to distilled/deion

ized water and bring volume to 10.0mL. Adjust pH to 8.0 with NaOH.

Mix thoroughly. Sparge with 100% N2. Autoclave for 15 min at 15 psi

pressure–121°C.

Solution E (Vitamin Solution):

Composition per liter:

Pyridoxine-HCl........................................................................10.0mg

Thiamine-HCl·2H2O..................................................................5.0mg

Riboflavin ..................................................................................5.0mg

Nicotinic acid.............................................................................5.0mg

D-Ca-pantothenate......................................................................5.0mg

p-Aminobenzoic acid.................................................................5.0mg

Lipoic acid .................................................................................5.0mg

Biotin .........................................................................................2.0mg

Folic acid....................................................................................2.0mg

Vitamin B12 ..............................................................................0.10mg

Solution E (Vitamin Solution): Add components to distilled/de

ionized water and bring volume to 1.0L. Mix thoroughly. Sparge with

100% N2. Autoclave for 15 min at 15 psi pressure–121°C.

Solution F:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.4g

Preparation of Solution F: Add Na2S·9H2O to distilled/deionized

water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%

N2. Autoclave for 15 min at 15 psi pressure–121°C.

Solution G:

Composition per 10.0mL:

Na-dithionite ............................................................................... 0.25g

Preparation of Solution G: Add Na-dithionite to distilled/deion

ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with

100% N2. Autoclave for 15 min at 15 psi pressure–121°C.

Solution H:

Composition per liter:

NaOH ............................................................................................ 0.5g

Na2WO4·2H2O ...........................................................................4.0mg

Na2SeO3·5H2O...........................................................................3.0mg

Preparation of Solution H: Add components to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly. Sparge with 100%

N2. Filter sterilize.

Preparation of Medium: Gently heat solution A and bring to boil

ing. Boil solution A for a few minutes. Cool to room temperature. Gas

with 80% N2 + 20% CO2 gas mixture to reach a pH below 6. Autoclave

for 15 min at 15 psi pressure–121°C. Cool to room temperature. Se

quentially add 1.0mL solution B, 100.0mL solution C, 10.0mL solu

tion D, 10.0mL solution E, 10.0mL solution F, 1.0mL solution G, and

1.0mL solution H. Distribute anaerobically under 80% N2 + 20% CO2

into appropriate vessels.

Use: For the cultivation of Holophaga foetida.

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